Danggui Shaoyaosan Regulates Autophagy via AMPK/mTOR/ULK1 Signaling Pathway in Rat Model of Metabolism-associated Fatty Liver Disease / 中国实验方剂学杂志

ObjectiveTo investigate the regulatory effect of Danggui Shaoyaosan on adenosine monophosphate-activated protein kinase （AMPK）/mammalian target of rapamycin （mTOR）/Unc-51-like kinase-1 （ULK1） signaling pathway in the rat model of metabolism-associated fatty liver disease （MAFLD）. MethodSixty SD rats were randomized into control， model， western medicine （polyene phosphatidylcholine capsules，0.144 g·kg-1）， and low-， medium-， and high-dose （2.44， 4.88， 9.76 g·kg-1， respectively） Danggui Shaoyaosan groups. After being fed with a high-fat diet for 8 weeks， the rats in each group were administrated with corresponding drugs for 4 weeks. At the end of drug treatment， serum and liver tissue were collected for subsequent determination of related indicators. ResultCompared with the control group， the model group showed increased contents of total cholesterol （TC）， triglyceride （TG）， low-density lipoprotein cholesterol （LDL-C） and activities of alanine aminotransferase （ALT） and aspartate aminotransferase （AST） in the serum， increased contents of TC， TG， and free fatty acids （FFAs） in the liver （P<0.01）， and decreased content of high-density lipoprotein cholesterol （HDL-C） in the serum （P<0.01）. Furthermore， the model group showed down-regulated protein levels of p-AMPK， microtubule-associated protein 1 light chain 3B （LC3B） Ⅱ， Beclin1， and ULK1 （P<0.01） and up-regulated protein levels of p-mTOR and ubiquitin-binding protein p62 in the liver （P<0.01）. The hepatic steatosis was obvious and the NAFLD activity score （NAS） and oil red O staining area increased in the model group， （P<0.05， P<0.01）. Compared with the model group， Danggui Shaoyaosan reduced the contents of TC and TG and the activities of ALT and AST in the serum， lowered the levels of TC， TG， and FFA in the liver， down-regulated the protein levels of p-mTOR and p62 （P<0.01）， elevated the serum HDL-C level， and up-regulated the protein levels of p-AMPK， LCBⅡ， Beclin1， and ULK1 in the liver （P<0.05， P<0.01）. Moreover， it alleviated hepatic steatosis and decreased the NAS and oil red O staining area （P<0.05， P<0.01）. ConclusionDanggui Shaoyaosan has therapeutic effect on MAFLD rats by regulating AMPK/mTOR/ULK1 signaling pathway to enhance autophagy.

Feasibility of Two-Screw Anterior Fixation for Odontoid Fractures in a Chinese Population:A Morphometric Study Based on Computed Tomography

Background@#To evaluate the feasibility of treating odontoid fractures in the Chinese population with two cortical screws based on computed tomography (CT) scans and describe a new measurement strategy to guide screw insertion in treating these fractures. @*Methods@#A retrospective review of cervical computed tomographic scans of 128 patients (aged 18–76 years; men, 55 [43.0%]) was performed. The minimum external transverse diameter (METD), minimum external anteroposterior diameter (MEAD), maximum screw length (MSL), and screw projection back angle (SPBA) of the odontoid process were measured on coronal and sagittal CT images. @*Results@#The mean values of METD and MEAD were 10.0 ± 1.1 mm and 12.0 ± 1.0 mm, respectively, in men and 9.2 ± 1.0 mm and 11.0 ± 1.0 mm, respectively, in women. Both measurements were significantly higher in men (p 9.0 mm that could accommodate two 3.5-mm cortical screws. The mean MSL value and SPBA range were 34.4 ± 2.9 mm and 13.5°–24.2°, respectively, with no statistically significant difference between men and women. @*Conclusions@#The insertion of two 3.5-mm cortical screws was possible for anterior fixation of odontoid fractures in 87 patients (68%) in our study, and there was a statistically significant difference between men and women.

Bioassay method for Compound Danshen Tablets based on a platelet aggregation inhibition test / 药学学报

A bioassay method for inhibiting platelet aggregation in vitro was established to quantify the pharmacological effects of Compound Danshen Tablets and support its quality control. The inhibition of platelet aggregation in rabbit plasma in vitro by Compound Danshen Tablets was used as the experimental system. The titer was calculated by using the method of dose-response parallel lines. As a result, a bioassay for the inhibition of platelet aggregation in vitro by Compound Danshen Tablets was established. Linearity was good in the concentration range of 0.128 g·mL-1 to 0.205 g·mL-1, and the titer of standard Compound Danshen tablets was 7 659 U·g-1 according to the titer definition. This in vitro assay was simple, reliable, reproducible and convenient. The activity of Compound Danshen Tablets in inhibiting platelet aggregation was quantified by potency assay and the quality of different batches was evaluated. The method can be applied for the quality control of Compound Danshen Tablets.

Network pharmacological study of Schizonepetae Herba and Saposhnikoviae Radix in treatment of ulcerative colitis / 中国中药杂志

The aim of this paper was to screen the active targets of Schizonepetae Herba and Saposhnikoviae Radix in the treatment of ulcerative colitis by means of network pharmacology,and to investigate their mechanism of action. The effective components of Schizonepetae Herba and Saposhnikoviae Radix were screened out by traditional Chinese medicine systematic pharmacological( TCMSP)database,with oral bioavilability( OB) ≥30% and drug-like( DL) ≥18% selected as the thresholds. Target PPI network was built between the main components and their corresponding targets. One hundred and eighty-two human genes corresponding to the medicine target sites were obtained from Uniprot database; 3 874 genes corresponding to ulcerative colitis were obtained from Genecard database.A total of 115 intersection genes were screened from disease genes and medicine genes,and the PPI interaction analysis was conducted by using String tool. Disease-target PPI network was drawn by using Cytoscape software,and component-target-disease network was constructed. One hundred and eight nodes and 1 882 connections were found,and then Cytoscape software was used to merge the networks and filter the core network for gene GO function analysis and KEGG pathway enrichment analysis. The mechanism of Schizonepetae Herba and Saposhnikoviae Radix was then verified by animal experiment. Gene GO functional analysis suggested that biological process,molecular functions and cell components were involved,and it was found that ulcerative colitis might be related to transcription factor activity,and cytokine receptor binding,etc. Gene KEGG pathway enrichment analysis showed that the mechanism of ulcerative colitis might be associated with TNF and Toll-like receptors( TLRs) signaling pathway-mediated cytoinflammatory factors interleukin-1( IL-1) and interleukin-6( IL6). The possible mechanism of the effective components of Schizonepetae Herba and Saposhnikoviae Radix in treating ulcerative colitis might be related to intervening the cytokine receptor binding of TNF and TLRs signaling pathways,reducing the transcription of nuclear factor-kappaB( NF-κB),and inhibiting the secretion of intestinal inflammatory factors IL-1 and IL-6.

Protective effect of DIZE on heart function of rats with diabetic cardio-myopathy / 中国病理生理杂志

AIM:To observed the protective effect of diminazene aceturate(DIZE),an angiotensin-converting enzyme 2(ACE2)activator,on rats with diabetic cardiomyopathy(DCM).METHODS:Male Wistar rats(n=30)were randomly divided into normal control group ,DCM group and DIZE treatment group(DIZE group).The rats in DCM group and DIZE group were intraperitoneally injected with streptozotocin(65 mg/kg )to establish diabetic model.After 12 weeks,the diabetic rats were infused with DIZE at 15 mg· kg-1 · d-1 or the same volume of saline for 4 weeks using os-motic minipump.The cardiac function was measured at the end of the 16th week.The methods of Mason staining and HE staining were used to observe the morphological changes of the myocardial tissue.Western blot ,ELISA and immunohisto-chemistry were used to observe the changes of ACE2,angiotensin(Ang)Ⅱ,Ang-(1-7),interleukin(IL)-1,IL-6 and connective tissue growth factor(CTGF).RESULTS:DIZE significantly improved the expression of ACE 2 in diabetic rats(P<0.05).Compared with DCM group,the levels of IL-1 and IL-6 in DIZE group were significantly decreased ,and the cardiac function in DIZE group was significantly improved(P<0.05).CONCLUSION:ACE2 endogenous agonist DIZE significantly increases the ACE 2 level and reduces the level of inflammation ,thus protecting the heart function of DCM rats.

Severe immune-related adverse events in the treatment with Pembrolizumab for a patient with ampullar carcinoma / 天津医药

Programmed death receptor-1 inhibitor pembrolizumab can restore the function of T cell activity and enhance the anti-tumor immune response by inhibiting the binding of PD-1 to its ligand PD-L1 and blocking the negative regulation of signal pathway. The activated T cells may cause immune-mediated adverse events in the process of anti-tumor. This article reported the severe immune related adverse effects induced by PD-1 inhibitor, pembrolizumab, in a patient with advanced ampullar carcinoma. The patient eventually died due to liver injury, leukocytosis,thrombocytopenia,and disseminated intravascular coagulation (DIC). This article reviewed the diagnosis and treatment of the patient, and reviewed the relevant literatures.

Flavanonol compounds from thorns of Gleditsie Spina and their cytotoxicity / 中草药

Objective: To study the flavonoids constituents from the thorns of Gleditsia sinensis and their cytotoxicity against tumor cells. Methods: The compounds were isolated by silica gel, Sephadex LH-20 column chromatography, and HPLC techniques. Their structures were elucidated on the basis of spectroscopic analyses. Results: Twelve flavonoids were obtained and identified as (2R,3R)-5,3',4'-trimethoxyl-7-hydroxyl-flavanonol (1), 5,7,3',4'-tetrahydroxyl-flavanonol (2), 5-methoxyl-3',4',7-trihydroxyl-flavanonol (3), dihydrokaempferol (4), epicatechin (5), 5,7,3',5'-tetrahydroxyl-flavanonol (6), fustin (7), (2R,3R)-7,3',5'-trihydroxyl-flavanonol (8), (2R,3R)-5,7,3'-trihydroxyl-4'-methoxyl-flavanonol (9), quercetin (10), 5,7,4'-trihydroxylflavone-8-C-glucopyranose (11), and 2,7-dimethyl-xanthone (12), respectively. Conclusion: Compound 1 is a new component named G. spina flavanonol A, and compounds 3,8,9, and 12 are isolated from the thorns of G. sinensis for the first time. The results of cytotoxicity test show that the dihydroflavonol compound 7 displays the stronger cytotoxicity against HepG2, A549, and EC109 cell strains, while compounds 1 and 3 have the effects on HepG2 and EC109, and compound 2 has the effect on EC109 cancer cells, respectively.

Design, synthesis and biological evaluation of novel 3-(2-oxo-2-substituted acetamido)benzamides as PARP-1 inhibitors / 药学学报

Poly(ADP-ribose)polymerase-1 (PARP-1) plays a significant role in the DNA repair process by catalyzing the transfer of ADP-ribose from NAD+ to its receptors. It is a promising anticancer drug target and many PARP-1 inhibitors have been developed and used in the clinical trial. In this work, a series of 3-(2-oxo-2-substituted acetamido)benzamides have been synthesized and their inhibitory activities against PARP-1 were evaluated. Of all the tested compounds, six compounds displayed inhibitory activities with IC50 values ranging from 0.23 to 5.78 µmol.L-1 . The binding pose of compound 5a was predicted using molecular docking to facilitate further structural modification.

Design, synthesis and biological evaluation of novel 3-(2-oxo-2-substituted acetamido)benzamides as PARP-1 inhibitors / 药学学报

Poly(ADP-ribose)polymerase-1 (PARP-1) plays a significant role in the DNA repair process by catalyzing the transfer of ADP-ribose from NAD+ to its receptors. It is a promising anticancer drug target and many PARP-1 inhibitors have been developed and used in the clinical trial. In this work, a series of 3-(2-oxo-2-substituted acetamido)benzamides have been synthesized and their inhibitory activities against PARP-1 were evaluated. Of all the tested compounds, six compounds displayed inhibitory activities with IC50 values ranging from 0.23 to 5.78 µmol.L-1 . The binding pose of compound 5a was predicted using molecular docking to facilitate further structural modification.

Computational chemistry in structure-based drug design / 药学学报

Today, the understanding of the sequence and structure of biologically relevant targets is growing rapidly and researchers from many disciplines, physics and computational science in particular, are making significant contributions to modern biology and drug discovery. However, it remains challenging to rationally design small molecular ligands with desired biological characteristics based on the structural information of the drug targets, which demands more accurate calculation of ligand binding free-energy. With the rapid advances in computer power and extensive efforts in algorithm development, physics-based computational chemistry approaches have played more important roles in structure-based drug design. Here we reviewed the newly developed computational chemistry methods in structure-based drug design as well as the elegant applications, including binding-site druggability assessment, large scale virtual screening of chemical database, and lead compound optimization. Importantly, here we address the current bottlenecks and propose practical solutions.

Computational chemistry in structure-based drug design / 药学学报

Today, the understanding of the sequence and structure of biologically relevant targets is growing rapidly and researchers from many disciplines, physics and computational science in particular, are making significant contributions to modern biology and drug discovery. However, it remains challenging to rationally design small molecular ligands with desired biological characteristics based on the structural information of the drug targets, which demands more accurate calculation of ligand binding free-energy. With the rapid advances in computer power and extensive efforts in algorithm development, physics-based computational chemistry approaches have played more important roles in structure-based drug design. Here we reviewed the newly developed computational chemistry methods in structure-based drug design as well as the elegant applications, including binding-site druggability assessment, large scale virtual screening of chemical database, and lead compound optimization. Importantly, here we address the current bottlenecks and propose practical solutions.

Correlation between changes of diffusion tensor imaging and clinical stages in patients with cerebral infarction / 中华神经医学杂志

Objective To evaluate the relationship between the changes of diffusion tensor imaging(DTI)and the clinical stages in patients with cerebral infarction. Methods One hundred and seven patients with cerebral infarction were chosen to perform conventional MRI and DTI,respectively.With double-blind method,the signal intensity of fractional anisotropy(FA)and isotropic apparent diffusion coefficient(ADCiso)in responsible to infraction regions was observed and based on their results,the relationships between the changes of DTI and the clinical stages were analyzed. Results In 29 cases of cerebral infarction in acute stage,10 were manifested as type I(hypointensity on ADCiso map,hyperintensity on FA map)and type Ⅱ(hypointensity on ADCiso map and FA map with hyper intensive circumference)on DTI;3 were manifested as type Ⅲ on DTI(iso-/Paypo-intensity on ADCiso map and FA map with normal circumference);6 were manifested as type Ⅳ on DTI(hyperintensity on ADCiso map,hypointensity on FA map).In 34 cases of cerebral infarction in sub-acute stage,2 were manifested astype Ⅰ on DTI;20 were manifested astype Ⅱ;8 were manifested as type Ⅲ and 4 were manifested as type IV.In 44 cases of cerebral infarction chronic stage.1 Was manifested as type Ⅱ on DTI;2 were manifested as type Ⅲ and 41 were manifested as type IV.Positive correlation was found between changes of DTI and clinical stages(r=0.693,P=0.000). Conclusion The intensity of infraction on DTI can directly demonstrate the clinical stages,providing precise evidence for classifying the stages of cerebral infarction.

Effect of anti-EGFR monoclonal antibody on chemosensitivity of human colon cancer cells and the mechanism / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of anti-EGFR monoclonal antibody on the chemosensitivity of human colon cancer cells and explore the possible molecular mechanism.</p><p><b>METHODS</b>The inhibitory effect of irinotecan (CPT-11), oxaliplatin (L-OHP) and fluorouracil (5-Fu), used alone or in combination with anti-EGFR monoclonal antibody, on the proliferation of LoVo cells in vitro was assessed by MTT assay. The expressions of PI3K and Akt protein in the treated cells were examined by Western blotting, and their mRNA expressions were detected by RT-PCR.</p><p><b>RESULTS</b>Both h-R3 and C-225 treatments significantly increased the chemosensitivity of LoVo cells to irinotecan and oxaliplatin. 5-Fu and h-R3 coadministered showed a synergistic effect on the cells, but 5-Fu and C-225 had an antagonistic action. Treatment with C-225 or h-R3 resulted in lowered expressions of PI3K and Akt in LoVo cells.</p><p><b>CONCLUSION</b>Anti-EGFR monoclonal antibody can increase the chemosensitivity of human colon cancer cells to most chemotherapeutic drugs, and such effect might be attributed to the blocking of PI3K/Akt signaling pathway by these antibodies.</p>

Clinical significance of VEGF levels in serums of colorectal cancer patients at stage IV / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the clinical significance of vascular endothelial growth factor (VEGF) levels in serums of colorectal cancer patients at stage IV.</p><p><b>METHODS</b>Using enzyme linked immunosorbent assay (ELISA) to detect the VEGF levels in serums of 45 colorectal cancer patients at stage IV, and 20 healthy served as normal control.</p><p><b>RESULTS</b>The mean concentration of VEGF in 45 colorectal cancer patients at the 7 day after operation were significantly lower than that before operation (P<0.01). The mean concentration of VEGF in the patients who benefit from bevacizumab showed no statistical difference from the levels of who did not benefit (P=0.554).</p><p><b>CONCLUSION</b>The VEGF levels in colorectal patients at stage IV are lowed as the load of tumor decrease. The circulating levels of VEGF seem not predict the response to bevacizumab in colorectal cancer patients at stage IV.</p>

Study on the peripheral blood dendritic cells subtypes and the expression of co-stimulating molecules on dendritic cells and B cells in severe aplastic anemia patients / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the quantities of monocyte-derived dendritic cell precursors (pDC1) and plasmacytoid dendritic cell precursors (pDC2) in peripheral blood mononuclear cells (PBMC) of severe aplastic anemia (SAA) patients before and after immune suppressive therapy (IST), the ratio of the pDC1 to pDC2, and the expression of co-stimulating molecules (CD80, CD86, CD40) on dendritic cells (DC) and B cells in SAA patients.</p><p><b>METHODS</b>By means of three color monoclonal antibody labeling technology, the quantities and ratio of pDC1 and pDC2 in PBMC were detected in 26 SAA patients at active phase, 13 at recovery phase and 15 normal controls respectively. The aforementioned parameters of 10 SAA patients were tested before and 2 months after IST. The expression of CD80, CD86 and CD40 on DC and B lymphocytes were detected in 16 SAA patients and 15 normal controls.</p><p><b>RESULTS</b>The percentages of pDC1 and the ratio of pDC1/pDC2 of controls were (0.41 +/- 0.05)% and 1.58 +/- 0.18 respectively, and those of SAA patients at active phase were (0.67 +/- 0.13)% and 2.70 +/- 0.32 respectively, [pDC1 (P < 0.05); pDC1/ pDC2 ratio (P < 0.01)]. The aforementioned parameters in convalescent SAA patients decreased to (0.43 +/- 0.10)%, and 1.78 +/- 0.36 respectively, being no difference from those of normal controls. The percentages of pDC1 and pDC2 in 10 SAA patients were (0.87 +/- 0.31)%, and (0.35 +/- 0.09)%, before IST, and (0.24 +/- 0.09)%, (0.14 +/- 0.04)%, after IST, being significantly decreased (P < 0.05). The percentages of CD86 expression on DC of controls was (11.97 +/- 4.31)%, and that of SAA patients was (29.84 +/- 3.02) % (P < 0.05). The percentages of CD80, CD40 and CD86 expression on lymphocytes of controls were (2.57 +/- 0.44)%, (7.34 +/- 1.22)% and (1.86 +/- 1.11)%, respectively, and those of SAA patients were (5.17 +/- 0.68)%, (8.85 +/- 2.94)% and (5.98 +/- 0.96)% respectively (P < 0.05, P < 0.01). The percentage of CD86 expression on B lymphocytes in controls was 8.04 +/- 0.66%, and in SAA patients was (20.46 +/- 2.78)%, (P < 0.05).</p><p><b>CONCLUSION</b>The pDC subtypes were abnormal and the percentage of pDC1 is increased in SAA patients, which are associated with stage of this disease. DC and B Lymphocytes in SAA patients upregulated expression of costimulatory molecules (CD86) which cause the T lymphocyte abnormally activated.</p>

Burden of abnormal hematopoietic clone in patients with myelodysplastic syndromes / 中国医学科学杂志(英文版)

<p><b>OBJECTIVE</b>To investigate the role of the burden of abnormal hematopoietic clone in the development of myelodysplastic syndromes (MDS).</p><p><b>METHODS</b>The ratio of the bone marrow cells with abnormal chromosomes to the total counted bone marrow cells was regarded as the index of MDS clone burden. The disease severity related parameters including white blood cell count, hemoglobin, platelet count, lactate dehydrogenase level, bone marrow blast, myeloid differentiation index, micromegakaryocyte, transfusion, interleukin-2, tumor necrosis factor (TNF), CD4+ and CD8+ T cells of MDS patients were assayed, and the correlations between those parameters and MDS clone burden were also analyzed.</p><p><b>RESULTS</b>The clone burden of MDS patients was 67.4% +/- 36.2%. MDS clone burden positively correlated with bone marrow blasts (r = 0.483, P < 0.05), negatively with hemoglobin level (r = -0.445, P < 0.05). The number of blasts, hemoglobin, and erythrocytes in high clone burden (> 50%) and low clone burden ( < or = 50%) groups were 7.78% +/- 5.51% and 3.45% +/- 3.34%, 56.06 +/- 14.28 g/L and 76.40 +/- 24.44 g/L, (1.82 +/- 0.48) x 10(12)/L and (2.32 +/- 0.66) x 10(12)/L, respectively (all P < 0.05). CD4+ T lymphocytes of MDS patients and normal controls were (0.274 +/- 0.719) x 10(9)/L and (0.455 +/- 0.206) x 10(9)/L, respectively (P < 0.05). CD8+ T lymphocytes of MDS patients and normal controls were (0.240 +/- 0.150) x 10(9)/L and (0.305 +/- 0.145) x 10(9)/L, respectively. The serum level of interleukin-2 of MDS patients (6.29 +/- 3.58 ng/mL) was significantly higher than normal control (3.11 +/- 1.40 ng/mL, P < 0.05). The serum level of TNF of MDS patients and normal control group were 2.42 +/- 1.79 ng/mL and 1.68 +/- 0.69 ng/mL, respectively. The ratio of CD4 to CD8 was higher in high clone burden MDS patients (1.90 +/- 0.52) than that in low clone burden patients (0.97 +/- 0.44, P < 0.05).</p><p><b>CONCLUSION</b>The quantitive clonal karyotype abnormalities and deficient T cell immunity are important parameters for evaluating MDS severity and predicting its progression.</p>

The prognostic value of quantitative chromosomal abnormality in myelodysplastic syndromes / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the prognostic value of quantitative chromosomal abnormality in myelodysplastic syndromes (MDS).</p><p><b>METHODS</b>Chromosomal karyotypes in seventy-one MDS patients' were analyzed quantitatively. Based on the number of abnormal metaphase in 20 counted metaphases, the patients were divided into three groups: no abnormal karyotypes, abnormal metaphases less than or equal to five, and that more than five. The leukemia transformation rate, death rate and survival time between these three groups were compared.</p><p><b>RESULTS</b>Forty-four cases (62.0%) had abnormal karyotypes. The incidences of abnormal karyotypes in RA, RCMD and RAEB were 76.9%, 55.8% and 75.0%, respectively, being no significant difference (P > 0.05). Among the abnormal karyotypes, complex abnormality with two or more abnormal karyotypes was most common and accounted for 47.7%. The frequencies of trisomy 8, monosomy 7 and del 20q were 18.2%, 4.5% and 4.5%, respectively. Other kinds of abnormal karyotypes totally accounted for 25%. There were 27 cases of group 1, 28 of group 2 and 16 of group 3. Eighteen cases (25.4%) transformed to acute leukemia. The incidences of leukemia transformation in group 1, 2 and 3 were 18.5%, 25% and 37.5%, and the death rates were 29.6%, 42.9% and 56.3%, respectively. The median survival times were 60, 47 and 24 months respectively.</p><p><b>CONCLUSION</b>The quantitative chromosome abnormality has prognostic value in MDS.</p>

Expression of apoptosis-related proteins in bone marrow neutrophils of patients with paroxysmal nocturnal hemoglobinuria / 中国实验血液学杂志

This study was aimed to evaluate expression levels of CD166, Fas and apoptosis-related proteins in bone marrow neutrophils of PNH patients and normal controls, and to analyze their correlation in order to explore whether exist apoptosis abnormality in BM neutrophils of PNH patients. The expression levels of CD16b, Fas and Bax, Bcl-2 in BM neutrophils of PNH patients and normal controls were assayed by flow cytometry; the difference of expression levels between patients and controls, and expression correlation between CD16b and apoptosis-related proteins were compared. The results showed that (1) the expression levels of CD16b on BM neutrophils of patients and controls were (20.36 +/- 9.05)% and (71.34 +/- 26.8)% respectively (P = 0.01); (2) the expression levels of CD95 on BM neutrophils of patients and controls were (62.83 +/- 32.11)% and (48.00 +/- 38.52)% respectively, there were no significant difference between CD95 expressions in BM neutrophils of PNH patients and controls and no significant correlation between expression of CD95 and CD16b on BM neutrophils of PNH patients (P > 0.05); (3) the expression levels of Bcl-2 in BM neutrophil cytoplasma of patients and controls were (8.64 +/- 5.40)% and (16.82 +/- 15.39)% respectively, there were no significant difference between Bcl-2 expression of patients and controls, and no significant correlation between the expression of Bcl-2 and CD16b in BM neutrophil cytoplasma of PNH patients (P > 0.05); (4) the expression levels of Bax in BM neutrophil cytoplasma of patients and control were (30.47 +/- 22.15)% and (48.47 +/- 15.99)% respectively, there were no significant difference between the Bax expressions of patients and controls, and no significant correlation between the Bax and CD16b expressions in BM neutrophil cytoplasma of PNH patients. In conclusion, BM neutrophils of PNH patients expressed apoptosis-related CD95, Bcl-2 and Bax without significant difference from the normal controls, and without significant correlation with the CD16b expression. It is suggested that the cell growth and decrease of PNH patients possibly are independent of abnormal apoptosis.

Expression of cell cycle control genes in myelodysplastic syndromes / 中华血液学杂志

<p><b>OBJECTIVE</b>To study aberrant expression of cell cycle control genes in patients with myelodysplastic syndromes (MDS).</p><p><b>METHODS</b>Reverse transcription polymerase chain reaction (RT-PCR) was used to investigate the expression of cell cycle control genes (cyclin D2, cyclin D3, cyclin A1, cyclin E, CDK2, CDK4, CDK6, p21, p27, p57, Rb and E2F1) in bone marrow mononuclear cells (BMMNCs) from 29 normal control, 27 MDS and 19 de novo acute myeloid leukemia (AML).</p><p><b>RESULTS</b>The expression levels of cyclin D3 (0.65 +/- 0.17, P < 0.05) and cyclin A1 (0.48 +/- 0.04, P < 0.05) in MDS were higher than those in normal control and significantly lower than those in AML. The expression rates and levels of cyclin D2 (40.7% and 0.78 +/- 0.21) and cyclin E (51.9% and 0.52 +/- 0.10) in MDS were statistically higher than those in normal control and AML. The expression level of CDK2 in MDS (0.66 +/- 0.19, P < 0.01) was higher than that in normal control (0.42 +/- 0.04) and the expression rate of CDK6 in MDS (25.9%) higher than in normal control (3.4%, P < 0.05). There was no significant difference of the expression rates and levels of CDK4 in MDS, AML and normal control. The expression rates and levels of p21 (77.8% and 1.18 +/- 0.21) and p27 (48.1% and 1.14 +/- 0.40) in MDS were statistically higher than those in normal control and AML. The expression level of p57 in MDS (0.69 +/- 0.06) was higher than that (0.53 +/- 0.05, P < 0.01) in normal control but lower than in AML (0.96 +/- 0.16, P < 0.01). The expression rate (55.6%) and level (0.85 +/- 0.17) of Rb in MDS were significantly higher than those in normal control and AML. The expression rate (7.4%) and level (0.39 +/- 0.04) of E2F1 in MDS were comparable to those in normal control but lower than those in AML.</p><p><b>CONCLUSION</b>MDS clones have aberrant mechanism of cell cycle control: high expressions of cyclin family members, CDK2 and CDK6 may lead to high proliferation; high expression of p21 and p27 may cause the G1 phase arrest.</p>

Study of Th cell subsets in bone marrow of myelodysplastic syndromes patients / 中华血液学杂志

<p><b>OBJECTIVE</b>To study the quantity and ratio of Th1, Th2 cells in the bone marrow of myelodysplastic syndromes (MDS) patients, and to evaluate the correlation between the ratio of the blast cells and the number of the Th1 cells in the bone marrow of MDS patients.</p><p><b>METHODS</b>By FACS, the quantity and ratio of IFN-gamma producing CD4(+) T cell (Th1) and IL-4 producing CD4(+) T cell (Th2) cells in the bone marrow were detected in 21 MDS patients, 18 normal controls and 13 severe aplastic anemia (SAA) patients respectively. The karyotypes of 18 MDS patients and 15 normal controls were assayed. The correlation between the ratio of the blast cells in the bone marrow and the number of the Th1 cells in the MDS patients were analyzed.</p><p><b>RESULTS</b>The percentages of Th1 cells, Th2 cells and ratio of Th1/Th2 in the bone marrow of normal controls were (0.48 +/- 0.10)%, (0.24 +/- 0.19)% and 2.31 +/- 0.76 respectively, while those of the MDS patients were (0.36 +/- 0.11)%, (0.76 +/- 0.35)% and 0.51 +/- 0.13. The percentage of Th1 cells of patients with MDS was reduced and the Th1/Th2 ratio was significantly lower than that of normal controls (P < 0.01). Those of the patients with SAA were (4.75 +/- 0.49)%, (0.40 +/- 0.28)% and 26.5 +/- 8.79 respectively, their Th1 cells and Th1/Th2 ratio were markedly higher than those of normal controls (P < 0.01). In all of the 15 normal controls the karyotypes were normal, but that of MDS patients was (50.00 +/- 0.10)%. The lower ratio of the Th1 cells in the bone marrow of the patients with MDS and the AML which progressed from MDS was negatively correlated with the higher percentage of the blast cells (r = -0.563, P < 0.01).</p><p><b>CONCLUSIONS</b>(1) The immune function of T lymphocytes in MDS is abnormal: the balance between Th1 and Th2 cells is broken. (2) With descending of the number of Th1 cells in the bone marrow of the MDS patients, the disease is progressing to leukemia.</p>